目的 建立青藤碱、雷公藤甲素皮肤和血液在体微透析方法。方法 以青藤碱和雷公藤甲素的回收率(R)为指标,采用增量法考察灌流液体积流量对R的影响,并综合考虑实际操作时需要注意的因素,确定青藤碱和雷公藤甲素微透析的体积流量和取样间隔;采用增量法和减量法考察灌流液质量浓度对探针体外R以及传递率(D)的影响,确定探针R和D之间的比例关系;用在体微透析减量法,考察一定时间间隔内皮肤和血液探针的D,并确定探针R的稳定性。结果 增量法中,青藤碱、雷公藤甲素的皮肤和血液探针体外R在0.5～2.5 μL/min,随着体积流量的增高而降低,综合考虑到实际实验时间和数据准确性,最后确定体积流量为1 μL/min,采样时间间隔为0.5 h;体积流量为1 μL/min、青藤碱质量浓度为10～40 μg/mL和雷公藤甲素质量浓度为3～12 μg/mL时,青藤碱、雷公藤甲素皮肤和血液探针体外R和D稳定且相等,说明药物质量浓度对微透析探针R影响小、在体微透析可以用探针D代替R;在体微透析减量法在10 h内测得的青藤碱和雷公藤甲素皮肤探针体内D分别为(41.27±0.87)%和(37.8±0.99)%,血液微透析探针体内D分别为(44.68±1.28)%和(51.35±1.15)%,说明青藤碱和雷公藤甲素在10 h内探针的D保持稳定。结论 建立的青藤碱和雷公藤甲素皮肤与血液微透析方法可用于青藤碱和雷公藤甲素皮肤给药后皮肤和血液药动学研究。

Objective To establish skin and blood microdialysis (MD) method of sinomenine and triptolide. Methods Incremental method was applied and the recovery rate of sinomenine and triptolide was used as an index to investigate the impact of perfusion fluid flow rate to the recovery (R). The flow rate and sampling interval were then determined according to the R and actual operation; The reduction method and incremental method were applied in the in vitro study to investigate impact of perfusate concentration to the R and a transfer rate (D) and the proportional relationship between them; Reduction method was applied in the in vivo study to investigate and determine the stability of the probe recoveries. Results The recovery of skin and blood probes in vitro of sinomenine and triptolide increased with the flow rate in the range of 0.5—2.5 μL/min. The flow rate was determined to be 1 μL/min and the sampling interval was determined to be 0.5 h; The R of sinomenine and triptolide in skin and blood probes in vitro and D was stable and equal despite of change of sinomenine concentration in 10—40 μg/mL and triptolide concentration in 3—12 μg/mL, indicating that the effect of drug concentration on the MD probe R was small and that R could be replaced by the D; The D in vivo using MD method was measured at 10 h. The D values of sinomenine skin and blood probe were (41.27 ± 0.87)% and (37.8 ± 0.99)%, respectively and those of triptolide were (44.68 ± 1.28)% and (51.35 ± 1.15)%, respectively, indicating that the D of sinomenine and triptolide was stable within 10 h. Conclusion The established method of sinomenine and triptolide in MD can be applied to investigate the kinetics in skin and blood after sc administration of formulation.

广东省教育部产学研结合项目(2012B091100486);中央财政支持地方高校发展专项资金 [276(2014)]